However, none of the reference genes discovered thus far is consistently expressed in a universal and invariant way under various experimental conditions, and recent reference gene selection studies indicate that a single reference gene is generally insufficient to normalize the expression data of all target genes. In insect, the reference genes have been validated at least in the desert locust Schistocerca gregaria, the oriental fruit fly Bactrocera dorsalis, the fruit fly Drosophila melanogaster emerald ash borer Agrilus planipennis, the diamondback moth Plutella xylostella, the tobacco whitefly Bemisia tabaci, and the red imported fire ant, Solenopsis invicta under a diverse set of biotic and abiotic conditions. However, no one single universal reference was identified, either. Therefore, it is not surprising that no single universal reference is available for four different lepidopteran insect species. In this context, reliable reference genes for gene expression analysis based on different experimental conditions should be selected. The tobacco whitefly, B. tabaci, is an invasive insect pest of agriculture and horticulture worldwide. Because of the application of chemical insecticides has been the MK-4827 primary strategy for the control of B. tabaci, this pest has developed different levels of resistance to a wide range of insecticides. It has been well documented that insecticide resistance in B. tabaci usually is associated with enhanced detoxification by oxidative and hydrolytic pathways. Therefore, increasing numbers of studies are using the RT-qPCR techniques to detect the changes of mRNA expression of detoxifying enzymes genes in resistant populations and tried to provide new insights into insecticides resistance mechanisms. Though the suitable references genes has been documented in bacterially challenged bees, in Tribolium beetles infected with fungus, in plant virus infected B. tabaci and in Bt toxin treated P. xylostella, this kind of information is lacking for insects stressed by different types of chemical insecticides. In this study, a set of reliable reference genes for gene expression analysis in the B. tabaci biotype Q, one of the most invasive and destructive pest in the world, after exposure to eight commonly used insecticides was selected and then valuated with two target genes, a P450 gene and glutathione S-transferase gene. Over expression of the P450 has been proved to be responsible for neonicotinoid insecticides resistance in B. tabaci while this is not the case for GST. The objective of this work is to provide a set of universal reliable reference genes for research of genes with toxicological function in B. tabaci. There exists no doubt that the qRT-PCR technology has made the quantitative determination of gene expression more convenient than ever before. However, extreme care must be taken for the selection of internal reference genes before their application of qRT-PCR. It has been documented that the selection of suitable reference genes is very important to obtain reliable and accurate data.